Investigation of Endophytic Bacterial Community Structure within the Tobacco Roots Using Culture-independent Techniques

doi:10.3969/j.issn.1000-7091.2012.01.038

Abstract

Abstract: To the aim of study was to investigate endophytic bacterial diversity in tobacco root, to lay a foundation for furthering to tap. We investigated endophytic bacterial diversity in tobacco root from Yunnan by culture-independent method. Total DNA genome of tobacco root sample was extracted using CTAB procedure with some modifications. A pair of bacterial PCR primers was used for endophytic bacterial 16 S rDNA amplification and a clone library was constructed for the tobacco root DNA samples. Clones screened from clone library on the basis of MspⅠ digestion patterns were sequenced and compared, flowed by constructing Neighbor-Joining tree. The results show that in total 155 clones were grouped to 36 operational taxonomic units, most of the clones showed high similarity to the known cultured bacteria. Sequence analysis revealed diverse phyla of bacteria in the 16 S rDNA library, which consisted of alpha, gamma, Beta subclasses of the Proteobacteria, Actinobacteria, Bacteroides, Firmicutes and Uncultured bacteria; 86. 2% of the clones were highly related to the known bacteria in the genus Enterobacter, Acinetobacter, Stenotrophomonas, Pseudomonas, Methylobacillus, Acidovorax, Bacillus etc; whereas 13. 5% of the clones showed higher affiliation with uncultured bacteria(93%-99% sequence similarity) and may represent novel taxa. The dominant bacterial groups of tobacco endophytic bacteria belong to Pseudomonas genera. There was abundant endophytic bacterial diversity in tobacco root from Yunnan, and there were large number of unknown endophytic bacterial groups here.

Key words: Culture-independent, Endophytic bacterial, Tobacco, Diversity of microorganism

CLC Number:

S432

CHEN Ze-bin, XIA Zhen-yuan, LEI Li-ping, LIU Fei, CHEN Hai-ru, ZHONG Yong-li. Investigation of Endophytic Bacterial Community Structure within the Tobacco Roots Using Culture-independent Techniques[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2012, 27(1): 201-209. doi: 10.3969/j.issn.1000-7091.2012.01.038.

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